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Escherichia coli remains the first choice of many researchers for producing recombinant proteins due to ease of use, rapid cell growth and low cost of culturing

Liquid-liquid extraction, either with acetonitrile based on QuEChERS (Quick, Easy, Cheap, Effective, Rugged, Safe) protocols or by salting-out assisted extraction with ethyl acetate are other commonly used sample preparation techniques used also for large cohorts (Njumbe Ediage et al

Consistent data on long-term use in children are limited, though adult studies report discontinuation rates between 37% and 81% at one year due to factors like cost, insurance, comorbidities, and absence of type 2 diabetes [83,84,85]

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